Application of chitosan/Fe₃O₄ microsphere-graphene composite modified carbon ionic liquid electrode for the electrochemical detection of the PCR product of soybean Lectin gene sequence.

In this paper a Fe(3)O(4) microsphere, graphene (GR) and chitosan (CTS) nanocomposite material modified carbon ionic liquid electrode (CILE) was used as the platform for the construction of a new electrochemical DNA biosensor. The single-stranded DNA (ssDNA) probe was immobilized directly on the surface of the CTS/Fe(3)O(4)-GR/CILE, which could hybridize with the target ssDNA sequence at the selected conditions. By using methylene blue (MB) as the electrochemical indicator the hybridization reaction was investigated with the reduction peak current measured. By combining the specific properties such as the biocompatibility and big surface area of Fe(3)O(4) microspheres, the excellent electron transfer ability of GR, the good film-forming ability of CTS and the high conductivity of CILE, the synergistic effects of nanocomposite increased the amounts of ssDNA adsorbed on the electrode surface and then resulted in the greatly increase of the electrochemical responses. Under the optimal conditions differential pulse voltammetric responses of MB were proportional to the specific ssDNA sequences concentration in the range from 1.0×10(-12) to 1.0×10(-6)mol/L with the detection limit as 3.59×10(-13)mol/L (3σ). This DNA biosensor showed good stability and discrimination ability to one-base and three-base mismatched ssDNA sequences. The polymerase chain reaction (PCR) product of soybean Lectin gene sequence was detected by the proposed method with satisfactory result, suggesting that the CTS/Fe(3)O(4)-GR/CILE was a suitable sensing platform for the sensitive detection of specific gene sequence.